Phosphatidylserine peroxidation/externalization during staurosporine-induced apoptosis in HL-60 cells
Phosphatidylserine peroxidation/externalization during staurosporine-induced apoptosis in HL-60 cells
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DOI：
10.1016/s0014-5793(02)02990-3
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发表时间：
2002-07-31
期刊：
影响因子：
3.5
通讯作者：
Kagan, VE
中科院分区：
文献类型：
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作者：
Matsura, T;Serinkan, BF;Kagan, VE
虽然氧化应激通常与细胞凋亡有关，但其在细胞凋亡程序执行中的具体作用尚未得到描述。我们假设，带负电荷的磷脂酰丝氨酸（PS）和带正电荷的细胞色素e之间的催化氧化还原相互作用释放到胞质溶胶中，沿着活性氧（ROS）的产生，导致PS的显著氧化和外化，以及随后的巨噬细胞对凋亡细胞的识别。通过使用staurosporine，蛋白激酶抑制剂，不作为促氧化剂，我们能够诱导HL-60细胞凋亡，而不触发非特异性氧化反应的混杂效应。通过这种方法，我们首次证明了PS在非氧化剂诱导的细胞凋亡的早期阶段（2小时）经历了统计学显著和显著的氧化，而最丰富的磷脂，磷脂酰胆碱，没有。谷氨酰胺（GSH），最丰富的细胞溶质硫醇，也保持未氧化在这个时间点。此外，PS氧化和细胞色素c在胞质溶胶中的外观是并行的; PS外化之后是凋亡细胞的吞噬作用。这些发现与我们提出的氧化PS依赖信号在细胞凋亡和吞噬作用的作用是兼容的。(C)2002年由Elsevier Science B. V.代表欧洲生物化学学会联合会出版。
Although oxidative stress is commonly associated with apoptosis, its specific role in the execution of the apoptotic program has yet to be described. We hypothesized that catalytic redox interactions between negatively charged phosphatidylserine (PS) and positively charged cytochrome e released into the cytosol, along with the production of reactive oxygen species (ROS), results in pronounced oxidation and externalization of PS, and subsequent recognition of apoptotic cells by macrophages. By using staurosporine, a protein kinase inhibitor that does not act as a prooxidant, we were able to induce apoptosis in HL-60 cells without triggering the confounding effects of nonspecific oxidation reactions. Through this approach, we demonstrated for the first time that PS underwent a statistically significant and pronounced oxidation at an early stage (2 h) of non-oxidant-induced apoptosis while the most abundant phospholipid, phosphatidylcholine, did not. Glutathione (GSH), the most abundant cytosolic thiol, also remained unoxidized at this time point. Furthermore, PS oxidation and the appearance of cytochrome c in the cytosol were concurrent; PS externalization was followed by phagocytosis of apoptotic cells. These findings are compatible with our proposed roles for oxidative PS-dependent signaling during apoptosis and phagocytosis. (C) 2002 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.